This tutorial shows the recording of FLIM-FRET images using an LSM upgrade kit, in this case connected to an Olympus FluoView FV1200 laser scanning microscope. In FLIM-FRET measurements, the fluorescence lifetime of a FRET-donor in presence or absence of the acceptor is measured. This can be used to monitor interactions between two molecules via FRET. The principles are similar as for any FLIM-recording. General recording of FLIM images can also be found using the link below.
The general principles of the data acquisition are analogous in all LSM upgrade kits. The movie was recorded in 02/2018; the recording procedure may be slightly different in older or newer models.